RESUMEN
Approximately 50 genes other than heat shock protein (HSP) expression changes during thermal stress. These genes like nitric oxide synthase (NOS) need proper attention and investigation to find out their possible role in the adaptation to thermal stress in animals. So, the present study was undertaken to demonstrate the expressions of inducible form type II NOS (iNOS), endothelial type III NOS (eNOS), constitutively expressed enzyme NOS (cNOS), HSP70, and HSP90 in peripheral blood mononuclear cells (PBMCs) during different seasons in Barbari goats. Real-time polymerase chain reaction, western blot, and immunocytochemistry were applied to investigate messenger RNA (mRNA) expression, protein expression, and immunolocalization of examined factors. The mRNA and protein expressions of iNOS, eNOS, cNOS, HSP70, and HSP90 were significantly higher (P < 0.05) during peak summer, and iNOS and eNOS expressions were also observed to be significantly higher (P < 0.05) during peak winter season as compared with moderate season. The iNOS, eNOS, cNOS, HSP70, and HSP90 were mainly localized in plasma membrane and cytoplasm of PBMCs. To conclude, data generated in the present study indicate the possible involvement of the NOS family genes in amelioration of thermal stress so as to maintain cellular integrity and homeostasis in goats.
Asunto(s)
Cabras/genética , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP90 de Choque Térmico/genética , Trastornos de Estrés por Calor/genética , Trastornos de Estrés por Calor/veterinaria , Óxido Nítrico Sintasa/genética , Animales , Femenino , Cabras/fisiología , Proteínas HSP70 de Choque Térmico/metabolismo , Proteínas HSP90 de Choque Térmico/metabolismo , Trastornos de Estrés por Calor/metabolismo , Leucocitos Mononucleares/metabolismo , Óxido Nítrico Sintasa/metabolismo , ARN Mensajero/metabolismo , Estaciones del AñoRESUMEN
Changing climatic scenario with expected global rise in surface temperature compelled more focus of research over decoding heat stress response mechanism of animals and mitigation of heat stress. Recently betaine, a trimethyl form of glycine has been found to ameliorate heat stress in some species of animals. To overcome deleterious effect of heat stress, an attempt was taken to investigate the effect of betaine supplementation on heat stress mitigation in goats. Eighteen female Barbari goats were taken and randomly divided into 3 groups (n=6) such as control, HS (Heat stressed), HS+B (Heat stressed administered with betaine). Except for the control group, other groups were exposed to repeated heat stress (42 °C) for 6 h for sixteen consecutive days. Blood samples were collected at the end of heat exposure on day 1 (Initial heat stress acclimation - IHSA), day 6 (Short term heat stress acclimation - STHSA) and day 16 (Long term heat stress acclimation - LTHSA). When the groups were compared between different heat stress acclimatory phases, expression of all HSPs (HSP60, HSP70, HSP90 and HSP105/110) showed a similar pattern with a first peak on IHSA, reaching a basal level on STHSA followed by second peak on LTHSA. The messenger RNA (mRNA) and protein expression of HSPs was observed to be higher (P<0.05) in HS group than HS+B group except HSP90 on IHSA and HSP60 on STHSA. HSP105/110 expression was highest (P<0.05) on LTHSA. Immunocytochemical analysis revealed that HSPs were mainly localized both in nucleus and cytoplasm of PBMCs. In conclusion, heat stress increases HSPs expression and betaine administration was shown to have a dwindling effect on expression of HSPs, suggesting a possible role of this chemical chaperone on heat stress amelioration.
Asunto(s)
Betaína/administración & dosificación , Cabras/fisiología , Proteínas de Choque Térmico/sangre , Proteínas de Choque Térmico/genética , Aclimatación , Animales , Betaína/farmacología , Núcleo Celular/metabolismo , Citoplasma/metabolismo , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Cabras/sangre , CalorRESUMEN
Menacing global rise in surface temperature compelled more focus of research over understanding heat stress response mechanism of animals and mitigation of heat stress. Twenty-four goats divided into four groups (n = 6) such as NHS (non-heat-stressed), HS (heat-stressed), HS + VC (heat-stressed administered with vitamin C), and HS + VE + Se (heat-stressed administered with vitamin E and selenium). Except NHS group, other groups were exposed to repeated heat stress (42 °C) for 6 h on 16 consecutive days. Blood samples were collected at the end of heat exposure on days 1, 6, 11, and 16. When groups compared between days, expression of all heat shock proteins (HSPs) showed a similar pattern as first peak on day 1, reached to basal level on the sixth day, and followed by second peak on day 16. The relative messenger RNA (mRNA) and protein expression of HSP 60, HSP70, and HSP90 was observed highest (P < 0.05) in HS group, followed by antioxidant-administered group on days 1 and 16, which signifies that antioxidants have dampening effect on HSP expression. HSP105/110 expression was highest (P < 0.05) on day 16. We conclude that HSP expression pattern is at least two-peak phenomenon, i.e., primary window of HSP protection on the first day followed by second window of protection on day 16. HSP60, HSP70, and HSP90 play an important role during the initial phase of heat stress acclimation whereas HSP105/110 joins this cascade at later phase. Antioxidants may possibly attenuate the HSP expression by reducing the oxidative stress.
Asunto(s)
Cabras/metabolismo , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/veterinaria , Proteínas de Choque Térmico/metabolismo , Adaptación Fisiológica , Animales , Cabras/fisiología , Proteínas de Choque Térmico/genética , Calor , Leucocitos Mononucleares/metabolismo , ARN Mensajero/metabolismoRESUMEN
Six, nonpregnant, Barbari goats aged 4-5 years were selected for the study. For the first 6 days, the animals were kept in psychrometric chamber at thermoneutral temperature for 6 h each day to make them acclimated to climatic chamber. On the 7th day, the animals were exposed to 41 °C temperature for 3 h and then to 45 °C for the next 3 h. Cardinal physiological responses were measured, and blood samples (3 ml) were collected at 1-h interval during the heat exposure period and then once after 6 h of the heat exposure. The rectal temperature (RT) and respiratory rate (RR) increased significantly (P < 0.05) during the heat exposure compared to pre- and postexposure. The relative messenger RNA (mRNA) expression of heat shock protein (HSP)60, HSP70, and HSP90 increased significantly (P < 0.05) within 1 h after exposure to heat stress at 41 and 45 °C and decreased significantly (P < 0.05) in next 2 h but remain significantly (P < 0.05) elevated from preexposure. HSP105/110 relative mRNA expression level remained unchanged during the first 4 h, and thereafter, it increased significantly (P < 0.05) and reached the peak at 6 h. Relative protein expression pattern of HSPs during exposure to heat stress showed similar trend as observed for the relative mRNA expression. Given the response sensitivity and intensity of HSP genes to environmental stresses, HSP70 was found to be the most sensitive to temperature fluctuation, and it could be used as an important molecular biomarker to heat stress in animals.
Asunto(s)
Cabras , Trastornos de Estrés por Calor/genética , Trastornos de Estrés por Calor/fisiopatología , Proteínas de Choque Térmico/genética , Animales , Temperatura Corporal , Cabras/genética , Cabras/metabolismo , Cabras/fisiología , Frecuencia Cardíaca , Trastornos de Estrés por Calor/metabolismo , Trastornos de Estrés por Calor/veterinaria , Proteínas de Choque Térmico/metabolismo , Humedad , Leucocitos Mononucleares/metabolismo , ARN Mensajero/metabolismo , Frecuencia Respiratoria , TemperaturaRESUMEN
Recent experiments using expression, immunolocalization, and cell culture approaches have provided leading insights into regulation of luteal angiogenesis by different growth factor systems and its role in the function of corpus luteum (CL) in buffalo. On the contrary, lymphangiogenesis and its regulation in the CL are still poorly understood. The aim of this study was to evaluate the expression and localization of lymphangiogenic factors (vascular endothelial growth factor [VEGF]-C and VEGFD), their receptor (VEGFR3), and lymphatic endothelial marker (LYVE1) in bubaline CL during different stages of the estrous cycle and to investigate functional role of VEGFC and VEGFD in luteal lymphangeogenesis. The mRNA and protein expression of VEGFC, VEGFD, and VEGFR3 was significantly greater in mid and late luteal phases, which correlated well with the expression of LYVE1. The lymphangiogenic factors were localized in luteal cells, exclusively in the cytoplasm. Immunoreactivity of VEGFC was greater during midluteal phase and that of VEGFD was greater during the mid and late luteal phases. Luteal cells were cultured in vitro and treated for different time duration (24, 48, and 72 hours) with VEGFC and VEGFD each at 50, 100, and 150 ng/mL concentration and VEGFC with VEGFD at 100 ng/mL concentration. The temporal increase in LYVE1 mRNA expression was significant (P < 0.05) in VEGFC and VEGFC with VEGFD treatment and no significant change was seen in VEGFD treatment. Thus, it seems likely that VEGFD itself has little role in lymphangiogenesis but along with VEGFC it might have a synergistic effect on VEGFR3 receptors for inducing lymphangiogenesis. In summary, the present study provided evidence that VEGFC and VEGFD, and their receptor VEGFR3, are expressed in bubaline CL and are localized exclusively in the cell cytoplasm, suggesting that these factors have a functional role in lymphangiogenesis of CL in buffalo.
Asunto(s)
Búfalos/metabolismo , Cuerpo Lúteo/metabolismo , Ciclo Estral/metabolismo , Receptores de Hialuranos/metabolismo , Linfangiogénesis , Factor A de Crecimiento Endotelial Vascular/metabolismo , Receptor 3 de Factores de Crecimiento Endotelial Vascular/metabolismo , Animales , Células Cultivadas , Cuerpo Lúteo/efectos de los fármacos , Femenino , InmunohistoquímicaRESUMEN
The present study has demonstrated the expression of HSP60, HSP70, HSP90, and UBQ in peripheral blood mononuclear cells (PBMCs) during different seasons in three different age groups (Groups I, II, and III with age of 0-2, 2-5, and >5 years, respectively) of goats of tropical and temperate regions. Real-time polymerase chain reaction was applied to investigate mRNA expression of examined factors. Specificity of the desired products was documented using analysis of the melting temperature and high-resolution gel electrophoresis to verify that the transcripts are of the exact molecular size predicted. The mRNA expression of HSP60, HSP90, and UBQ was significantly higher (P < 0.05) in all age groups during peak summer season as compared with peak winter season in both tropical and temperate region goats. HSP70 mRNA expression was significantly higher (P < 0.05) during summer season as compared with winter season in tropical region goats. However, in the temperate region, in goats from all the three age groups studied, a non-significant difference of HSP70 expression between summer and winter seasons was noticed. In conclusion, results demonstrate that (1) HSP genes are expressed in caprine PBMCs and (2) higher expression of HSPs during thermal stress suggest possible involvement of them to ameliorate deleterious effect of thermal stress so as to maintain cellular integrity and homeostasis in goats.