Quantitative patterns of expression of gap junction genes during in vivo or in vitro development of ovarian follicles in sheep

Influence of TCM 199B, α-MEM, Waymouth MB 752/1 culture media, VEGF, Estradiol-17β, GDF-9 and FGF on in vitro development of preantral follicles in sheep.

“…Day on which the PFs’ were placed in culture was designated day zero and subsequent days as 1, 2, 3 and so on. Half of the medium was replaced by an equal volume of fresh medium every 48 h. This culture procedure supported good development of sheep PFs’ in earlier studies ( Chakravarthi et al., 2015 , 2016a , 2016b ; Kamalamma et al., 2016 ; Kona et al., 2016 ; Kumar et al., 2019 ; Lakshminarayana et al., 2014 ; Srividya et al., 2017 ). Each follicle was morphologically evaluated every 24 h during culture period using an inverted microscope (Leica, DMIRB, Germany) for the proportions of follicles exhibiting growth, increase in the diameter and antrum formation ( Fig.…”

“…Bicarbonate buffered tissue culture medium 199 (TCM 199B) was supplemented with 50 µg/mL gentamicin sulfate, 1 µg/mL l -thyroxine (T 4 ), 2.0 µg/mL follicle stimulating hormone (FSH), 10 ng/mL insulin like growth factor-I (IGF-I) and 1MIU/mL of growth hormone (GH). Unlike in our previous studies ( Arunakumari et al., 2010 , 2007 ; Chakravarthi et al., 2015 , 2016a , 2016b ; Hemamalini et al., 2003 ; Kamalamma et al., 2016 ; Kona et al., 2016 ; Kumar et al., 2019 ; Lakshminarayana et al., 2014 ; Rajarajan et al., 2006 ; Srividya et al., 2017 ; Tamilmani et al., 2005 ), FSH (F8174) and LH (L5269) used in the present study were of ovine rather than porcine and equine origin respectively. This medium referred to as the standard medium in the present study, routinely supports good development of sheep PFs’ in vitro ( Arunakumari et al., 2010 , 2007 ; Chakravarthi et al., 2015 , 2016a , 2016b ; Hemamalini et al., 2003 ; Kamalamma et al., 2016 ; Kona et al., 2016 ; Kumar et al., 2019 ; Lakshminarayana et al., 2014 ; Rajarajan et al., 2006 ; Srividya et al., 2017 ; Tamilmani et al., 2005 ).…”

“…Barring the media supplemented with different growth factors and hormones, all the other solutions were filtered through a 0.22 µm sterilizing filter (Sartorius, Germany) prior to use. All the procedures employed in the study were previously reported from the laboratory ( Arunakumari et al., 2010 , Arunakumari et al., 2007 ; Chakravarthi et al., 2015 , Chakravarthi et al., 2016a , Chakravarthi et al., 2016b ; Hemamalini et al., 2003 ; Kamalamma et al., 2016 ; Kona et al., 2016; Kumar et al., 2019; Lakshminarayana et al., 2014; Rajarajan et al., 2006 ; Srividya et al., 2017 ;; Tamilmani et al., 2005 ;). However, a sufficiently brief explanation of all the methods employed in this study is presented.…”

Granulosa cell genes that regulate ovarian follicle development beyond the antral stage: the role of estrogen receptor β

“…Moreover, a sharp decrease of selective transcript levels after hCG administration suggests that either LH/LHCGR signaling leads to abrupt downregulation of transcription or degradation of those mRNAs. Such spatiotemporal patterns of expression in response to gonadotropins are characteristic of many of the GC genes involved in follicle maturation (Bédard et al, 2003;Chakravarthi et al, 2015;Chakravarthi et al, 2016;Chen et al, 2009;Lakshminarayana et al, 2014;Ronen-Fuhrmann et al, 1998). However, those reports were based on studying one or a few genes, but our study demonstrates the spatiotemporal pattern of gene expression among the whole transcriptome in wildtype and Erβ null GCs.…”

Advances in in vitro folliculogenesis in domestic ruminants

“…Despite all these advances on IVFC of secondary follicles, oocyte maturation and embryo production rates are still far below the results obtained from follicles entirely grown in vivo. On this regard, several studies with this follicular category have shown that IVFC (isolated follicles in a 2D system) negatively affect the expression pattern and/or level of genes related to oocyte survival and development such as P450 aromatase (Lakshminarayana et al, 2014); B-cell leukemia/lymphoma -2 (Bcl2) and Bcl2-associated X protein (Bax) (Praveen Chakravarthi et al, 2015); connexins 32 and 43 (CX32 and CX43) (Chakravarthi et al, 2016a); cyclin B1 (CCNB1) and cyclin D1 (CND1) (Chakravarthi et al, 2016b); and GDF-9 and BMP-15 (Kona et al, 2016).…”