Thumbnail Image

Theses (Ph.D.)

Browse

2018 - 201922020 - 20221
 Reset filters

Settings

Subject: Biochemistry ×

Search Results

Now showing 1 - 3 of 3
  • Thumbnail Image
    ThesisItemOpen Access
    ELUCIDATION OF CELLULAR AND MOLECULAR EFFECTS OF HERBAL PLANT EXTRACTS AND THEIR BIOACTIVE COMPOUNDS ON MURINE AND OVINE OVARIAN FUNCTIONS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR, 2022) B. SAMPATH KUMAR; V. GIRISH KUMAR
    The relevance of plants and/or plant extracts in the field of animal reproduction has been undeniable since the dawn of time. Proper understanding and behavior of phytochemicals on both in vivo and in vitro models are required to employ them in the field of animal reproduction to augment fertility. As a result, the present study aimed to determine the cellular and molecular effects of herbal plant extracts (Murraya Koenigii: MK, Moringa Oleifera: MO, Aegle Marmelos: AM, Mimosa pudica: MP) and their bioactive compounds (Carbazole, Kaempferol, Quercetin, Coumarin, and Mimosine) on murine and ovine ovarian functions. The above mentioned plant leaves were extracted with methanol and given orally to cyclical mature 8-week rats at various concentrations (100, 300, and 1000mg/kg b.wt.) for three consecutive estrous cycles. MK and AM leaf extracts were able to increase ovarian follicular population at 1000mg/kg, but MO extracts only did so at 300mg/kg. The MP extracts, on the other hand, had a detrimental influence on follicular growth. In a second experiment, quercetin and coumarin compounds were given to a different set of rats, both orally and parenterally, at 50mg/kg and 25mg/kg b.wt. for three cycles. Quercetin was effective on follicle growth even at 25mg/kg in both routes. Coumarin, on the other hand, was only efficacious in the parenteral route at 50mg/kg. In rats, however, quercetin was even more effective than coumarin in terms of ovarian follicle development. The effects of the aforementioned bioactive substances on sheep ovarian functions were studied by culturing them with isolated granulosa cells and preantral follicles. When cultured with carbazole, kaempferol, quercetin, myricetin, and coumarin, 25μM,10μM,10μM,25μM and 10μM concentrations respectively were effective for granulosa cell development and 50μM,10μM,25μM,25μM and 25μM concentrations respectively were effective for preantral follicle growth. Mimosine, on 341 the other hand, had a detrimental influence on GC and PF development in vitro. Gene expression studies were also conducted to corroborate the beneficial effects of the aforesaid substances on GCs and PFs. When cultured with carbazole (25, 50μM), kaempferol (both 10μM), quercetin (10, 25μM), myricetin (both 25μM), and coumarin (10, 25μM) at selective concentrations for GCs and PFs respectively, FGF2, GDF9, CYP19, and BCL2 were expressed nearly 2 fold higher in the GCs as well as PFs when compared to the expression of GAPDH. However, BAX expression was 2fold lower. Similarly, even at 10 μM, BAX expression was increased by more than 2.5 times when cultured with mimosine. Further proteomic analysis of partially purified spent media proteins of cultured GCs and PFs on MALDI –TOF-MS identified a Prohibitin II (33.29KDa) in the Fraction G (85-95%) of 10μM quercetin+GC cultures and CYP7A1 (56.88 KDa) in the Fraction G of 25μM quercetin+ PF cultures spent media. Serum albumin was identified in the Fraction F (75-85%) of 10μM coumarin + GC cultured spent media.
  • Thumbnail Image
    ThesisItemOpen Access
    CELLULAR AND GENOMIC STUDIES ON OVINE OOCYTE-GRANULOSA CELL INTERACTION AND APOPTOSIS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR-585 401, 2018-07-01) RAMESH, H. S.; V. GIRISH KUMAR
    The experiments were designed to examine the ovine ovarian follicular population and investigate the effect of different concentrations of AREG, NRG-I and TNF-α at 5ng, 10ng, 25ng, 50ng, 100ng and 150ng and their best dose combinations on growth of PFs, oocyte of PFs, COCs maturation, COCs expansion, cleavage rate, morula/blastocyst formation, GCs/CCs monolayer formation, gene expression and secretory protein of invitro cultured LAFs, MAFs and SAFs. In the first experiment, there was significant changes (P<0.05) in follicular population noticed during different seasons, different age Groups and at different follicular phases. In second experiment, there was significant increase in growth of PFs, oocyte, CCs expansion, maturation and granulosa monolayer formation observed at 100ng or 150ng AREG/NRG-I, 5ng or 10ng TNF-α, AREG+NRGI and AREG+NRG-I+TNF-α. In this experiment, COCs from LAFs at 50ng or 100ng or 150ng AREG/NRG-I and 25ng TNF-α, COCs from MAFs at 100ng or 150ng AREG, 50ng or 100ng or 150ng NRG-I and 25ng TNF-α, COCs from SAFs at 100ng or 150ng AREG, 150ng NRG-I and 5ng or 10ng TNF-α and COCs of LAFs, MAFs and SAFs cultured with AREG+NRG-I and AREG+NRG-I+TNF-α showed better cumulus expansion, maturation of oocytes, cleavage rate, morula or blastocyst formation and CCs monolayer formation. In third experiment, there was higher expression of caspase-3 at 150ng TNF-α, caspase-9 at 50ng of AREG, GDF-9 at 150ng AREG or AREG+NRG-I and IGF-I in 150ng of AREG, 25ng of TNF-α, AREG+NRG-I and AREG+NRG-I+TNF- α. In fourth experiment, oocytes retrieved from SPFs and LPFs revealed no secretory protein as evaluated by SDS-PAGE. Keywords: Preantral follicles, COCs, monolayer, AREG, NRG-I, TNF-α
  • Thumbnail Image
    ThesisItemOpen Access
    IMPACT OF HEAT STRESS ON DIFFERENT TOLL LIKE RECEPTORS EXPRESSION PATTERNS IN TWO DIFFERENT INDIGENOUS GOAT BREEDS
    (KARNATAKA VETERINARY, ANIMAL AND FISHERIES SCIENCES UNIVERSITY, BIDAR-585 401, 2019-07-01) SAVITHA, S. T.; V. GIRISH KUMAR
    The study was conducted to establish the differences in expression pattern of different toll-like-receptors (TLRs) in Osmanabadi and Salem Black goats and to compare the expression patterns of these TLRs between these breeds subjected to summer season induced heat stress. The animals were randomly allocated into four groups of six animals each, OC (n=6; Osmanabadi control); OHS (n=6; Osmanabadi heat stress); SC (n=6; Salem Black control) and SBHS (n=6; Salem Black heat stress). The OC and SC animals were maintained in the shed in comfort condition while OHS and SBHS animals were exposed outside to summer heat stress between 10:00h to 16:00h during experimental period. The results indicated that the expression of TLR1, TLR4, TLR5, TLR6 and TLR10 genes in both the breeds showed different expression pattern. The expression pattern of all these genes did not differ between the control (OC) and heat stress (OHS) group in Osmanabadi breed but significantly higher (P<0.05) expression of these genes were reported in heat stress group (SBHS) as compared to control group (SBC) in Salem Black breed. Further, the results indicated that the expression of TLR3 and TLR7 followed the same pattern between both Osmanabadi and Salem Black breeds with significantly higher (P<0.01) expression in heat stress group of both these breeds. However, TLR8 and TLR9 genes showed different expression patterns between the breeds with significantly higher (P<0.05) expression of these two genes recorded in Salem Black breed. The TLR2 expression was significantly higher (P<0.01) in heat stress group of Osmanabadi breed. But TLR2 did not expressed in Salem Black breed. The higher expression of most of TLRs in the SBHS group as compared to OHS group indicates the superior resilient capacity of Salem Black goats to maintain immune status even during exposure to adverse environmental condition. Keywords: Heat stress; Goat; Immune response; TLR; THI; Thermo-tolerance