RESUMO
In this study, we compared the quality of DNA extracted using the modified CTAB method, from Rheum palmatum leaves preserved using fourteen different methods, including ones used commonly in other species: under ultra-cold (-80°C) temperatures, after drying with an absorbent paper, desiccating using a silica gel, drying at 60°C, in 70% ethanol, absolute ethanol, 70% ethanol supplemented with 50 mM EDTA, SDS-DNA extracting solution, nuclear separation buffer, improved NaCl-CTAB solution, TE-buffer, I-solution, or II-solution. DNA extracted from fresh leaves was used as the control. The quality of extracted DNA was evaluated based on the success of PCR amplification of the ITS2 region and a microsatellite marker. DNA was not extracted from samples preserved in the nuclear separation buffer and II-solution. The purities of DNA extracted from leaves preserved in ultra-cold temperatures, 70% ethanol, and 70% ethanol with 50 mM EDTA, and after desiccating using a silica gel and drying were higher, and comparable to the purity of DNA extracted from fresh leaves, than those of leaves preserved using other methods. In the present study, combined with the PCR amplifications, the preservation using ultra-cold temperatures, silica gel desiccation, or drying, and PCR amplification of the extracted DNA can be used for further molecular studies in R. palmatum.
Assuntos
DNA de Plantas/isolamento & purificação , Preservação Biológica/métodos , Rheum/genética , Repetições de Microssatélites , Folhas de Planta/química , Folhas de Planta/genética , Reação em Cadeia da Polimerase/métodos , Rheum/químicaRESUMO
Congenital cataract is a common cause of blindness in children; however, its pathogenesis remains unclear. Genetic factors have been shown to play an important role in the pathogenesis of congenital cataract. The current genetic models of congenital cataract include autosomal dominant, autosomal recessive, and sex-linked inheritance. Sex-linked congenital cataract could be inherited through the X or Y chromosome. Congenital cataract is a symptom associated with several X-linked disorders, including Nance-Horan syndrome, Lowe syndrome, Conradi-Hünermann-Happle syndrome, oculo-facio-cardio-dental syndrome, and Alport syndrome. On the other hand, the mechanism and characteristics of Y-linked congenital cataract remains to be identified. Despite its rarity, sex-linked congenital cataract has been known to seriously affect the quality of life of patients. In this review, we present our current understanding of the genes and loci associated with sex-linked congenital cataract. This could help identify novel approaches for the prevention, early diagnosis, and comprehensive disease treatment.
Assuntos
Catarata/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Análise Mutacional de DNA , Genes Ligados ao Cromossomo X , Estudos de Associação Genética , Loci Gênicos , Predisposição Genética para Doença , Humanos , MutaçãoRESUMO
OBJECTIVE: The study aimed to evaluate the influence of repeated recruitment manoeuvres (RRMs) on lung injury in patients with acute respiratory distress syndrome (ARDS). METHODS: Forty-one ventilated patients with severe ARDS were selected for this study. Recruitment manoeuvres (RMs) were conducted with continuous positive airway pressure (CPAP; 30 cm H2O for 40 seconds). Recruitment manoeuvres were repeated every two hours for all three groups. Changes in haemodynamics, pulmonary compliance, gas exchange and extravascular lung water index (EVLWI) were monitored before RM (pre-RM), 10 minutes after each RM, and four hours after RM3 (4 hours post-RRM). Pulmonary inflammatory factors (tumour necrosis factor-alpha [TNF-α] and interleukin [IL]-6 and -10) were also analysed. RESULTS: Compared with those in pre-RM, pulmonary compliance, oxygenation index (ratio of partial pressure of arterial oxygen to fraction of inspired oxygen [PaO2/FiO2]) and EVLWI remarkably improved in RM1, RM2, RM3 and 4 hours post-RRM (p < 0.05). The PaO2/FiO2 ratio increased significantly in RM1 and RM3 (p < 0.05). Extravascular lung water index decreased significantly in RM1 compared with that in RM3 and 4 hours post-RRM (p < 0.05). There was no significant difference in cytokines. CONCLUSION: Repeated recruitment manoeuvres during lung-protected ventilation can improve pulmonary compliance and oxygenation and significantly decrease extravascular lung water in ARDS patients. Lung injury was not worsened by RRMs in patients with severe ARDS.
RESUMO
The use of noninvasive genetic sampling to identify the sex of wild animals is an extremely valuable and important tool in molecular ecology and wildlife conservation. Sex determination using the amelogenin gene has been conducted in many species because only a single pair of primers is required to amplify both X- and Y-linked alleles. However, this method has not been used in field research with the feces of wildlife. In this study, we applied this method to 222 fecal samples from wild blue sheep (Pseudois nayaur) using amelogenin primers (SE47/SE48) after testing the effectiveness of sex determination using tissue samples and fecal samples from blue sheep of known sex. We found this method to be highly reliable (80.2%) for blue sheep. Amelogenin can be used to identify the sex of wild animals using fecal samples.
Assuntos
Amelogenina/genética , Análise para Determinação do Sexo , Animais , Fezes , Análise de Sequência de DNA , Ovinos/genéticaRESUMO
The purpose of this study was to measure the genetic diversity of wild beach plum and cultivated species, and to determine the species relationships using SSRs markers. An analysis of genetic diversity from ten beach plum germplasms was carried out using 11 simple sequence repeat (SSR) primers selected from 35 primers to generate distinct PCR products. From this plant material, 44 allele variations were detected, with 3-5 alleles identified from each primer. The analysis showed that the genetic similarity coefficient varied from 0.721 ± 0.155 to 0.848 ± 0.136 within each of the ten beach plum germplasms and changed within the range of 0.551 ± 0.084 to 0.695 ± 0.073 between any two pairs of germplasms. According to the genetic dissimilarity coefficient matrix, a cluster analysis of SSRs using the unweighted pair group mean average method in the NTSYSpc 2.10 software revealed that the ten germplasms could be divided into two groups at the dissimilarity coefficient of 0.606. Class I included 77.8, 12.5, 30, and 33.3% of MM, MI, NY, and CM, respectively. Class II contains the remaining 9 beach plum germplasms. The markers generated by 11 SSR primers proved very effective in distinguishing the beach plum germplasm resources. It was clear that the geographical distribution did not correspond with the genetic relationships among the different beach plum strains. This result will be of value to beach plum breeding programs.
Assuntos
Variação Genética , Repetições de Microssatélites , Prunus domestica/genética , Análise por Conglomerados , Evolução Molecular , Filogenia , Polimorfismo Genético , Prunus domestica/classificaçãoRESUMO
We evaluated the effect of puerarin on spatial learning and memory ability of mice with chronic alcohol poisoning. A total of 30 male C57BL/6 mice were randomly divided into model, puerarin, and control groups (n=10 each). The model group received 60% (v/v) ethanol by intragastric administration followed by intraperitoneal injection of normal saline 30 min later. The puerarin group received intragastric 60% ethanol followed by intraperitoneal puerarin 30 min later, and the control group received intragastric saline followed by intraperitoneal saline. Six weeks after treatment, the Morris water maze and Tru Scan behavioral tests and immunofluorescence staining of cerebral cortex and hippocampal neurons (by Neu-N) and microglia (by Ib1) were conducted. Glutamic acid (Glu) and gamma amino butyric acid (GABA) in the cortex and hippocampus were assayed by high-performance liquid chromatography (HPLC), and tumor necrosis factor (TNF)-α and interleukin (IL)-1β were determined by ELISA. Compared with mice in the control group, escape latency and distance were prolonged, and spontaneous movement distance was shortened (P<0.05) by puerarin. The number of microglia was increased in both the cortex and hippocampal dentate gyrus (P<0.01), and neurons were reduced only in the hippocampal dentate gyrus (P<0.01) in puerarin-treated mice. In the model group, Glu and GABA levels decreased (P<0.05), and Glu/GABA, TNF-α, and IL-1β increased (P<0.01) with puerarin treatment, returning to near normal levels. In conclusion, puerarin protected against the effects of chronic alcohol poisoning on spatial learning and memory ability primarily because of anti-inflammatory activity and regulation of the balance of Glu and GABA.
Assuntos
Animais , Masculino , Etanol/intoxicação , Isoflavonas/uso terapêutico , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/prevenção & controle , Fármacos Neuroprotetores/uso terapêutico , Memória Espacial/efeitos dos fármacos , Vasodilatadores/uso terapêutico , Alcoolismo/complicações , Cromatografia Líquida de Alta Pressão , Córtex Cerebral/química , Córtex Cerebral/efeitos dos fármacos , Ensaio de Imunoadsorção Enzimática , Ácido Glutâmico/análise , Interleucina-1beta/análise , Isoflavonas/farmacologia , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/tratamento farmacológico , Microglia/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Distribuição Aleatória , Fatores de Tempo , Resultado do Tratamento , Fator de Necrose Tumoral alfa/análise , Vasodilatadores/farmacologia , Ácido gama-Aminobutírico/análiseRESUMO
This study investigated the effects of angiotensin II (AngII) intervention, using captopril and losartan, on the expression of matrix metalloproteinase-2 (MMP-2), MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1), and collagen in rats with pulmonary hypertension, in an effort to understand mechanisms underlying pulmonary vascular remodeling. A total of 40 male Sprague-Dawley rats were randomly divided into normal group, model group, captopril group, and losartan group. After 5 weeks, the mean pulmonary arterial pressure (mPAP), right ventricular index, and neointima formation in each group were determined. Immunohistochemical analysis was performed to determine the degree of pulmonary arterial muscularization as well as MMP-2, MMP-9, and TIMP-1 protein expression in lung tissue. Real-time fluorescent quantitative PCR was used to detect MMP2, MMP9, TIMP1, COL1A1, and COL4A1 mRNA expression. Picro-sirius red staining was performed to detect collagen protein expression. Neointima formation was observed in the model group. Moreover, the mPAP, right ventricular index, degree of arterial muscularization, and collagen deposition, as well as mRNA and protein expression of MMP2, MMP9, and TIMP1 were significantly higher than those in the other groups (P < 0.05). The mPAP, right ventricular index, degree of arterial muscularization, and mRNA and protein expression in the captopril and losartan groups were significantly decreased compared with those of the model group (P < 0.05). AngII regulates MMP-2, MMP-9, and TIMP-1 expression and affects collagen deposition. Thus, this hormone is involved in pulmonary vascular remodeling, indicating a possible mechanism that can be targeted in pulmonary hypertension intervention.
Assuntos
Angiotensina II/farmacologia , Colágeno Tipo IV/metabolismo , Colágeno Tipo I/metabolismo , Hipertensão Pulmonar/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Animais , Colágeno Tipo I/genética , Cadeia alfa 1 do Colágeno Tipo I , Colágeno Tipo IV/genética , Modelos Animais de Doenças , Hipertensão Pulmonar/tratamento farmacológico , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Artéria Pulmonar/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Inibidor Tecidual de Metaloproteinase-1/genéticaRESUMO
We evaluated the effect of puerarin on spatial learning and memory ability of mice with chronic alcohol poisoning. A total of 30 male C57BL/6 mice were randomly divided into model, puerarin, and control groups (n=10 each). The model group received 60% (v/v) ethanol by intragastric administration followed by intraperitoneal injection of normal saline 30 min later. The puerarin group received intragastric 60% ethanol followed by intraperitoneal puerarin 30 min later, and the control group received intragastric saline followed by intraperitoneal saline. Six weeks after treatment, the Morris water maze and Tru Scan behavioral tests and immunofluorescence staining of cerebral cortex and hippocampal neurons (by Neu-N) and microglia (by Ib1) were conducted. Glutamic acid (Glu) and gamma amino butyric acid (GABA) in the cortex and hippocampus were assayed by high-performance liquid chromatography (HPLC), and tumor necrosis factor (TNF)-α and interleukin (IL)-1ß were determined by ELISA. Compared with mice in the control group, escape latency and distance were prolonged, and spontaneous movement distance was shortened (P<0.05) by puerarin. The number of microglia was increased in both the cortex and hippocampal dentate gyrus (P<0.01), and neurons were reduced only in the hippocampal dentate gyrus (P<0.01) in puerarin-treated mice. In the model group, Glu and GABA levels decreased (P<0.05), and Glu/GABA, TNF-α, and IL-1ß increased (P<0.01) with puerarin treatment, returning to near normal levels. In conclusion, puerarin protected against the effects of chronic alcohol poisoning on spatial learning and memory ability primarily because of anti-inflammatory activity and regulation of the balance of Glu and GABA.
Assuntos
Etanol/intoxicação , Isoflavonas/uso terapêutico , Aprendizagem em Labirinto/efeitos dos fármacos , Transtornos da Memória/prevenção & controle , Fármacos Neuroprotetores/uso terapêutico , Memória Espacial/efeitos dos fármacos , Vasodilatadores/uso terapêutico , Alcoolismo/complicações , Animais , Córtex Cerebral/química , Córtex Cerebral/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Ensaio de Imunoadsorção Enzimática , Ácido Glutâmico/análise , Interleucina-1beta/análise , Isoflavonas/farmacologia , Masculino , Transtornos da Memória/induzido quimicamente , Transtornos da Memória/tratamento farmacológico , Camundongos Endogâmicos C57BL , Microglia/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Distribuição Aleatória , Fatores de Tempo , Resultado do Tratamento , Fator de Necrose Tumoral alfa/análise , Vasodilatadores/farmacologia , Ácido gama-Aminobutírico/análiseRESUMO
Rheum palmatum, one of the source plants of the traditional Chinese medicine rhubarb, is anendemic and endangered species. To our knowledge, this is the first report on the chromosome number and karyotype of this species. Sectioning combined with micrography was used to analyze the karyotype. The following results were obtained: R. palmatum had a stable chromosome number 2n = 22; the basic number of chromosomes was 11; karyotype formula is 2n = 22 = 20 metacentric + 2 submetacentric, belonging to Stebbins' 1A type; and karyotype asymmetry index was 55.39%. The present study showed that R. palmatum has a primitive type of karyotype.
Assuntos
Cromossomos de Plantas/genética , Espécies em Perigo de Extinção , Plantas Medicinais/genética , Rheum/genética , Cariótipo , Cariotipagem/métodos , Medicina Tradicional ChinesaRESUMO
Chloroplast microsatellite primers were developed in order to provide more population genetic information of endangered Rheum officinale, R. palmatum, and R. tanguticum for conservation. The dried roots and rhizomes of these plants are important in traditional Chinese medicine. The results showed that the optimum concentrations of Mg(2+), Taq DNA polymerase, dNTPs, template DNA, and primers in a 25-µL reaction system were 2.0 mM, 1.0 U, 0.10 mM, 20 ng, and 0.8 µM, respectively. Fourteen of 53 primer combinations were chosen for their high clarity and repetition in three species, and their annealing temperatures ranged from 56 to 58°C. These primers and the optimized polymerase chain reaction system may provide a tool for understanding the demography and genetic variation of these endangered plants.
Assuntos
DNA de Cloroplastos , Espécies em Perigo de Extinção , Repetições de Microssatélites , Rheum/genética , Primers do DNA , Reação em Cadeia da Polimerase/métodosRESUMO
This study aims to investigate the association of peroxisome proliferator-activated receptor (PPAR) delta -87T/C polymorphism with several sugar metabolism indices and tumor necrosis factor α (TNF α) level. The body mass index (BMI), waist size, and levels of fasting plasma glucose, serum lipid, fasting insulin, TNFα, and PPAR delta -87T/C of 286 patients with type 2 diabetes mellitus (T2DM) and 158 subjects with normal fasting glucose (NFG) were measured in a Dalian population. The distribution of genotypic frequencies between T2DM and NFG were not significantly different (χ(2) = 0.012, P = 0.994). BMI, fasting blood glucose (FBG), homeostasis model assessment-estimated insulin resistance (HOMA-IR), triglyceride, and TNFα levels were significantly different among different T2DM genotypes. HOMA-IR and FBG were significantly different among different NFG genotypes. The PPAR delta -87T/C polymorphism is known to be closely related with glucose levels and lipid metabolism. A close relationship was also found between HOMA-IR and TNFα levels and HOMA-IR and FBG in T2DM and NFG, respectively.
Assuntos
Diabetes Mellitus Tipo 2/genética , Obesidade/genética , PPAR delta/genética , Polimorfismo de Nucleotídeo Único , Regiões Promotoras Genéticas , Fator de Necrose Tumoral alfa/genética , Idoso , Alelos , Glicemia/metabolismo , Índice de Massa Corporal , Estudos de Casos e Controles , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/patologia , Jejum , Feminino , Frequência do Gene , Genótipo , Humanos , Insulina/sangue , Resistência à Insulina , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Obesidade/metabolismo , Obesidade/patologia , PPAR delta/metabolismo , Triglicerídeos/sangue , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Overexpression of cytokine-induced apoptosis inhibitor 1 (CIAPIN1) contributes to multidrug resistance (MDR) in breast cancer. This study aimed to evaluate the potential of CIAPIN1 gene silencing by RNA interference (RNAi) as a treatment for drug-resistant breast cancer and to investigate the effect of CIAPIN1 on the drug resistance of breast cancer in vivo. We used lentivirus-vector-based RNAi to knock down CIAPIN1 in nude mice bearing MDR breast cancer tumors and found that lentivirus-vector-mediated silencing of CIAPIN1 could efficiently and significantly inhibit tumor growth when combined with chemotherapy in vivo. Furthermore, Western blot analysis showed that both CIAPIN1 and P-glycoprotein expression were efficiently downregulated, and P53 was upregulated, after RNAi. Therefore, we concluded that lentivirus-vector-mediated RNAi targeting of CIAPIN1 is a potential approach to reverse MDR of breast cancer. In addition, CIAPIN1 may participate in MDR of breast cancer by regulating P-glycoprotein and P53 expression.
Assuntos
Animais , Feminino , Humanos , Antibióticos Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/genética , Inativação Gênica , Peptídeos e Proteínas de Sinalização Intracelular/genética , Western Blotting , Neoplasias da Mama/genética , Carcinoma/tratamento farmacológico , Carcinoma/genética , Modelos Animais de Doenças , Genes MDR , Vetores Genéticos/genética , Inibidores do Crescimento/genética , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lentivirus/genética , Camundongos Endogâmicos BALB C , Camundongos Nus , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/efeitos dos fármacos , Interferência de RNA , RNA Interferente Pequeno/genética , /efeitos dos fármacosRESUMO
This study assessed the clinical efficacy of lamivudine and adefovir dipivoxil combined with autologous bone marrow stem cell transplantation as treatment for patients with hepatitis B and decompensated liver cirrhosis. In total, 77 patients with hepatitis B and decompensated liver cirrhosis were randomly divided into two groups. Under general symptomatic and supportive treatment, the patients in group A (37 cases) were treated with lamivudine and adefovir dipivoxil, whereas those in group B (40 cases) were treated with autologous bone marrow stem cell transplantation in combination with lamivudine and adefovir dipivoxil. After 4 weeks of treatment, the liver function indicators and clinical signs and symptoms of the patients in group B improved more significantly than those of patients in group A. Lamivudine and adefovir dipivoxil in combination with autologous bone marrow stem cell transplantation effectively prevented hepatitis B virus infection and bone marrow stem cell damage. This combination treatment facilitates the differentiation of bone marrow stem cells into normal liver cells to restore liver structure and improve liver function, thereby improving the quality of life of patients.
Assuntos
Adenina/análogos & derivados , Vírus da Hepatite B/efeitos dos fármacos , Lamivudina/uso terapêutico , Cirrose Hepática/tratamento farmacológico , Cirrose Hepática/virologia , Organofosfonatos/uso terapêutico , Transplante de Células-Tronco , Adenina/efeitos adversos , Adenina/farmacologia , Adenina/uso terapêutico , Colinesterases/metabolismo , Terapia Combinada , Feminino , Humanos , Lamivudina/efeitos adversos , Lamivudina/farmacologia , Cirrose Hepática/fisiopatologia , Testes de Função Hepática , Masculino , Pessoa de Meia-Idade , Organofosfonatos/efeitos adversos , Organofosfonatos/farmacologia , Tempo de Protrombina , Transplante de Células-Tronco/efeitos adversos , alfa-Fetoproteínas/metabolismoRESUMO
Overexpression of cytokine-induced apoptosis inhibitor 1 (CIAPIN1) contributes to multidrug resistance (MDR) in breast cancer. This study aimed to evaluate the potential of CIAPIN1 gene silencing by RNA interference (RNAi) as a treatment for drug-resistant breast cancer and to investigate the effect of CIAPIN1 on the drug resistance of breast cancer in vivo. We used lentivirus-vector-based RNAi to knock down CIAPIN1 in nude mice bearing MDR breast cancer tumors and found that lentivirus-vector-mediated silencing of CIAPIN1 could efficiently and significantly inhibit tumor growth when combined with chemotherapy in vivo. Furthermore, Western blot analysis showed that both CIAPIN1 and P-glycoprotein expression were efficiently downregulated, and P53 was upregulated, after RNAi. Therefore, we concluded that lentivirus-vector-mediated RNAi targeting of CIAPIN1 is a potential approach to reverse MDR of breast cancer. In addition, CIAPIN1 may participate in MDR of breast cancer by regulating P-glycoprotein and P53 expression.
Assuntos
Antibióticos Antineoplásicos/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Doxorrubicina/uso terapêutico , Resistencia a Medicamentos Antineoplásicos/genética , Inativação Gênica , Peptídeos e Proteínas de Sinalização Intracelular/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/efeitos dos fármacos , Animais , Western Blotting , Neoplasias da Mama/genética , Carcinoma/tratamento farmacológico , Carcinoma/genética , Modelos Animais de Doenças , Feminino , Genes MDR , Vetores Genéticos/genética , Inibidores do Crescimento/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lentivirus/genética , Células MCF-7 , Camundongos Endogâmicos BALB C , Camundongos Nus , Interferência de RNA , RNA Interferente Pequeno/genética , Proteína Supressora de Tumor p53/efeitos dos fármacosRESUMO
The Brazilian short-tailed opossum, Monodelphis domestica, is born 14-15 days after copulation and is available for experimentation at stages of development corresponding to those which occur in utero in placental mammals. In the present study, we took advantage of the opossum's embryology to study the development of projections from caudal levels of the spinal cord to the brainstem and cerebellum using axonal tracing methods. In all cases, a 2-3 day survival time was used for axonal transport. When injections of Fast blue (FB) were made into caudal levels of the thoracic cord at postnatal day (PD) 1 or 2, axonal labeling could not be identified at supraspinal levels. When injections were made at PD3, however, labeled axons were found in the fasciculus gracilis at caudal medullary levels, within the ventrolateral medulla and pons, within an incipient inferior cerebellar peduncle, and within the cerebellar anlage. The dorsal root origin of at least some of the axons within the fasciculus gracilis was evidenced by the transganglionic transport of cholera toxin conjugated to horseradish peroxidase from the hindlimbs. After FB injections at PD7, a few labeled axons could be traced from the fasciculus gracilis into the nucleus gracilis and from the ventrolateral pathway to the inferior olive. Generally comparable results were obtained using wheat germ agglutinin conjugated to horseradish peroxidase (WGA-HRP). In cases injected with FB at PD9, the pattern of brainstem labeling was adult-like. Although labeled axons were present within the cerebellum of animals injected with FB on PD3, they were limited to the marginal zone. Axonal labeling was present within an identifiable internal granular layer in cases injected with either FB or WGA-HRP at PD16, and it appeared to be limited to specific bands which foreshadowed those seen at later stages of development and in the adult animal. In some cases, labeled axons were present within the molecular layer where they were not seen in the adult animal. Our results provide a timetable for the normal development of projections from caudal levels of the spinal cord to the brainstem and cerebellum in Monodelphis and show that such development occurs postnatally rather than prenatally, as in placental mammals.
Assuntos
Tronco Encefálico/crescimento & desenvolvimento , Cerebelo/crescimento & desenvolvimento , Gambás/crescimento & desenvolvimento , Medula Espinal/crescimento & desenvolvimento , Envelhecimento/fisiologia , Amidinas , Animais , Animais Recém-Nascidos , Transporte Axonal , Axônios/ultraestrutura , Tronco Encefálico/anatomia & histologia , Tronco Encefálico/citologia , Cerebelo/anatomia & histologia , Cerebelo/citologia , Corantes Fluorescentes , Peroxidase do Rábano Silvestre , Medula Espinal/anatomia & histologia , Medula Espinal/citologia , Conjugado Aglutinina do Germe de Trigo-Peroxidase do Rábano Silvestre , Aglutininas do Germe de TrigoRESUMO
We have used the retrograde transport of Fast blue (FB) to study the origins of supraspinal projections to the lumbar and cervical spinal cord at different stages of development in the Brazilian, short-tailed opossum, Monodelphis domestica. Monodelphis was chosen for study because its young are born in a very immature state, 14-15 days after copulation, making it possible to manipulate its nervous system in an embryonic state without intra-uterine surgery. When injections of FB were made into the lumbar cord at postnatal day (PD) 1, neurons were labeled within several areas of the reticular formation (the retroambiguus nucleus, the ventral and dorsal reticular nuclei of the medulla, the gigantocellular reticular nucleus, the lateral paragigantocellular reticular nucleus, and the pontine reticular nucleus), the presumptive coeruleus complex, and the lateral vestibular nucleus. In many cases, labeled neurons were also found within the caudal raphe and the presumptive interstitial nucleus of the medial longitudinal fasciculus. The results of immunocytochemical studies provided evidence for catecholaminergic and serotoninergic neurons in the brainstem at PD1 and for axons of both phenotypes in the spinal cord. By PD3, labeled neurons were found within the ventral gigantocellular and ventral pontine nuclei of the reticular formation, the spinal trigeminal nucleus, and the presumptive paraventricular nucleus of the hypothalamus. When injections were made at PD4, neurons were also labeled within the medial and inferior vestibular nuclei, the red nucleus, the mesencephalic nucleus of the trigeminal nerve, the presumptive nucleus of Edinger-Westphal and the lateral hypothalamus. By at least PD7, the pattern of supraspinal labeling was similar to that obtained at older ages and in the adult animal. When FB was injected into the cervical cord at PD1, neurons were labeled in all of the areas labeled by lumbar injections at the same age and in larger numbers. In addition, labeled neurons were found within the ventral gigantocellular and spinal trigeminal nuclei. When cervical injections were made at PD15, labeled neurons were found within the deep cerebellar nuclei and amygdala and by PD17 they were also present within the superior colliculus and cerebral cortex. In some cases, cortical labeling was present outside the areas labeled by comparable injections in adult animals.(ABSTRACT TRUNCATED AT 400 WORDS)