Characterization of salivary protein during ovulatory phase of menstrual cycle through MALDI-TOF/MS.

CONTEXT: Predicting ovulation is the basis on which the fertile period is determined. Nowadays there are many methods available to detect the ovulatory period. Unfortunately, these methods are not always effective for accurate detection of ovulation. Hence, an attempt was made to detect ovulation through single dimension sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of protein with the help of saliva ferning. AIMS: The aim of this study was to determine the association of protein level with endogenous reproductive hormone level across the menstrual cycle. SETTINGS AND DESIGN: Salivary protein and its confirmation were evaluated during menstrual cycle followed by SDS-PAGE and Mass spectrometry. STATISTICAL METHOD USED: The protein content present in saliva throughout menstrual cycle is trail by SPSS statistical software version. MATERIALS AND METHODS: Salivary proteins were investigated serially during pre-ovulatory, ovulatory and post-ovulatory periods of normal menstrual cycle in eighteen healthy volunteers. The samples were collected in three consecutive menstrual cycles. Salivary protein was estimated and analyzed by single dimension SDS-PAGE. RESULTS: The results revealed significant variations in protein concentrations during the menstrual cycle. Protein levels were maximum during ovulation and minimum during postovulatory phase. Further, single dimension SDS-PAGE analysis showed seven different fractions of proteins is from 14-90 kilo Dalton (kDa) in the three phases of the menstrual cycle. CONCLUSIONS: Among the proteins, 48 kDa protein was more predominantly exhibited during ovulatory phase than pre and post-ovulatory phase. The present study indicates that the protein level and the specific protein band (48 kDa) through MALDI-TOF MS analysis might serve as an indicator for ovulation.

The role of oxytocin release in the paraventricular nucleus in the control of maternal behaviour in the sheep.

Oxytocin (OT) release within the brain is thought to play a major role in inducing maternal behaviour in a number of mammalian species but little is known about the sites of release which are important in this respect. We have investigated whether the paraventricular nucleus of the hypothalamus (PVN) is a site of OT action on maternal behaviour in the sheep. In vivo microdialysis and retrodialysis was used to determine whether OT is released in the region of the PVN during the post-partum induction of maternal behaviour and if its release at this site can stimulate maternal behaviour in non-pregnant animals. In vivo sampling showed that OT concentrations increased significantly in the region of PVN at birth. When OT was retrodialysed bilaterally into the PVN (1 or 10 microM) of multiparous ewes treated with progesterone and oestradiol to stimulate lactation, maternal behaviour was induced in a significant number of animals (1 microM, 6/8 and 10 microM, 5/8) compared with controls (0/8 ewes). Similar infusions of the ring structure of OT, tocinoic acid (TOC-10 microM), also induced maternal behaviour in a significant proportion of animals (5/6 ewes) as did intracerebroventricular (ICV) OT (6/8 ewes) and artificial stimulation of the vagina and cervix (VCS, 8/9 ewes). On the other hand, vasopressin (AVP) 1 microM did not induce maternal behaviour in any ewes and a 10 microM dose only induced it in 2/8 animals. The neurochemical changes accompanying the above treatments were also investigated. Noradrenaline concentrations increased in the PVN after the retrodialysis administration of OT 1 microM and 10 microM, TOC 10 microM and AVP 1 microM, OT ICV and VCS. Dopamine concentrations were also increased by OT 10 microM, TOC 10 microM, AVP 1microM and OT ICV. Aspartate and glutamate concentrations were significantly reduced by retrodialysis infusions of OT 1 microM and AVP 1 and 10 microM but not by any other treatment. Finally, the retrodialysis infusion of OT and TOC, as well as ICV OT, significantly increased plasma OT release whereas AVP infusions did not. These results provide evidence that OT is released in the PVN during parturition and is important for the induction of maternal behaviour. It seems probable that OT release at this site has a positive feedback effect on both parvocellular and magnocellular OT neurons to facilitate co-ordinated OT release both in central OT terminal regions (to facilitate maternal behaviour) and peripherally into the blood (to facilitate uterine contractions/milk let down). The potential functional roles for the actions of OT on monoamine and amino acid transmitter release in the PVN are discussed.

Effect of substance P on acetylcholine and dopamine release in the rat striatum: a microdialysis study.

In vivo microdialysis in urethane anaesthetised rats was used to investigate the effects of substance P (SP) on acetylcholine (ACh) and dopamine (DA) release in the rat striatum. Results showed that SP elicited a dose-dependent increase in ACh release between 1 and 50 pmol/l. The rise in ACh release occurred both during SP administration and for up to 60 min after it. Dose-response curves either based on the initial rise in ACh release, or the total duration of increased release, showed a bell shape with 100 fmol/l and 5 nmol/l doses failing to significantly alter release and a 500 pM dose being less effective than 50 pmol/l. In contrast to this, SP did not significantly alter DA release at doses ranging between 100 fmol/l and 5 nmol/l. There was evidence for a strong desensitisation effect of SP administration since after initial treatment with SP subsequent doses of the peptide, even at very high doses, failed to provoke further changes in ACh still showed the expected increase in release in response to a potassium challenge. Physalaemin and neurokinin A increased ACh release with a similar potency to SP at a 50 pmol/l dose whereas neurokinin B and neuropeptide gamma, while increasing ACh release at a 50 pmol/l dose, were less potent than SP. The effect of SP on ACh release is probably mediated via NK-1 receptors since ACh release in response to SP was reduced in a dose dependent manner by the NK-1 receptor antagonists CP-96,345 and RP-67580.