Apoptosis-mediated testicular alteration in Japanese quail (Coturnix coturnix japonica) in response to temporal phase relation of serotonergic and dopaminergic oscillations

Abstract

Reproductive performance of many avian species, including Japanese quail, is reported to be modulated by specific temporal phase relation of serotonergic and dopaminergic oscillations. Accordingly, it has been shown that the serotonin precursor 5-HTP and the dopamine precursor L-DOPA given 8 h apart induce gonadal suppression and given 12 h apart lead to gonadal stimulation, while other temporal relationships were found to be ineffective. In the present study, we investigated the effects of 8-and 12-h phase relation of neural oscillations on testicular responses including expression of GnRH-I, GnIH, pro-apoptotic proteins ( p53 and Bax), inactive and active executioner caspase-3, and the uncleaved DNA repair enzyme PARP-1. Testicular volume and mass decreased significantly in 8-h quail and increased in 12-h quail compared with controls. Expression of ir-GnIH, p53, Bax and active-caspase-3 increased and that of GnRH-I, pro-caspase-3 and uncleaved PARP-1 decreased in 8-h quail compared with controls. A TUNEL assay also confirmed testicular regression in these quail. Testes of 12-h quail exhibited significantly increased expression of GnRH-I, pro-caspase-3 and uncleaved PARP-1 compared with the control group. Our findings suggest that differential response of avian testes to 8-and 12-h phase relation of serotonergic and dopaminergic neural oscillations may be attributed to autocrine/paracrine action of GnIH expression, which is upregulated in regressed testes, leading to apoptotic changes, and downregulated in developed testes, causing apoptotic inhibition. It is concluded that specific phase relation of neural oscillations may modulate the local testicular GnRH-GnIH system and alter the apoptotic mechanism in quail testes. Moreover, these findings highlight the physiological effects of time-dependent drug delivery, including the specific time intervals between two drugs.

Specific neural phase relation of serotonin and dopamine modulate the testicular activity in Japanese quail

“…The intensity of the bands was quantitated by gel densitometry (1.39-fold increase; p < 0.001) as compared with control (F (2,33) = 265.23; Figure 1). The intratesticular testosterone pattern is same as seen for the serum testosterone levels in our previous study (Banerjee et al, 2016). 4.4 | Testicular Cnx-43 and 14-3-3 immunoreactivity decreased in 8-hr quail while increased in 12-hr quail…”

2.45 GHz microwave radiation induced oxidative and nitrosative stress mediated testicular apoptosis: Involvement of a p53 dependent bax‐caspase‐3 mediated pathway

“…Immunofluorescence of full length p53 (rabbit polyclonal, sc‐6243, F‐393, 1:50), Bax (rabbit polyclonal, sc‐6236, 1:50), Bcl‐2 (mouse monoclonal, sc‐7382, 1:50), Bcl‐x L (mouse monoclonal, sc‐8392, 1:50), pro‐caspase‐3 (mouse monoclonal, sc‐7272, 1:50), cleaved caspase‐3 (rabbit monoclonal, MAB835, R&D Systems, Inc., Minneapolis, MN, 1:100), and uncleaved/full length PARP‐1 (mouse monoclonal, sc‐8007, 1:50) was carried out on neutral formalin‐fixed paraffin‐embedded testes sections using single and/or double immuno‐labeling/staining method. Immunofluorescence was performed in a two‐step procedure as described by Banerjee et al Briefly, in the first step, after initial deparafinization in xylene and rehydration in graded series of alcohol of testis section slides, antigen retrieval was performed using 1 M citrate buffer (pH 6) in a microwave oven at 1,000 W (3–4 min). Testes sections were then treated with blocking solution for 2 hr at room temperature [5% heat inactivated goat serum (HIGS)], and then sections were incubated with aforementioned mouse monoclonal and rabbit monoclonal/polyclonal antibodies for 24 hr in humid chamber.…”

Pueraria tuberosa extract inhibits iNOS and IL-6 through suppression of PKC-α and NF-kB pathway in diabetes-induced nephropathy

“…Immunofluorescence of PKC‐α, pNF‐kB, TNF‐α, IL‐6 and iNOS in kidney sections of all groups (normal control, the DN control and DN+PTY‐2r) was observed under a Zeiss LSM510 Meta laser‐scanning confocal microscope with a Plan‐Apo 20.0×, 1.4‐ NA oil immersion objective, and the images were collected using LSM 510 Meta software in same magnification and stored in TIFF files. Integrated optical density (IOD) of all images was measured and analysed using Image J software (Image J 1.48, National Institute of Health, Bethesda, MA, USA), and signal intensity of immune‐positive signals was measured as mentioned previously . The IOD values (arbitrary unit) were averaged (four sections per kidney and six kidneys per group) to determine the signal density.…”

“…Integrated optical density (IOD) of all images was measured and analysed using Image J software (Image J 1.48, National Institute of Health, Bethesda, MA, USA), and signal intensity of immune-positive signals was measured as mentioned previously. [35] The IOD values (arbitrary unit) were averaged (four sections per kidney and six kidneys per group) to determine the signal density. Based on the signal intensity, the term intense, moderate and weak immunoreactive cells were applied.…”