The effect of chronic kisspeptin

administration on seminal fructose levels

in male mice
Endocrine (2014) 45:144147
Amin Jan
Muhammad Haris Ramzan

Introduction
The kisspeptins have been identified as novel peptides having ability
to stimulate GnRH/LH/FSH secretion, through the activation of
GPR54, a classical G protein coupled receptor.
Kisspeptins, the product of KISS-1 gene, are neuropeptides belonging
to the family of RF-amide peptides.
They are encoded as a 145-amino acid product, Kp-145, that is
cleaved proteolytically into shorter peptides (Kp-54, -10, -13 and -14).
Kp-54 and its cleavage products share a common RF-amide Cterminal decapeptide and exhibit the same affinity and efficacy for
GPR54

Introduction
Fructose is secreted from the seminal vesicles and the accessory
sex glands.
It is the major carbohydrate found in seminal plasma and
appears essential for normal sperm motility.
The determination of fructose itself is of particular significance,
because there is a direct relationship between the fructose level
in sperm plasma and the testosterone function of the interstitial
cells of Leydig.
Low levels of Fructose may be the consequence of inflammation
in the prostrate or seminal vesicles, or structural abnormality of
the seminal vesicles and their ducts

Introduction
Seminal vesicles are androgen dependent.
It has been suggested that corrected seminal fructose may be used as a
biological marker of androgen activity in the reproductive tract.
In recent studies it has been demonstrated that continuous kisspeptin
exposure causes degeneration of seminal vesicle tissue as evidenced by
increased tubular lumen and decrease in epithelial height and epithelial
folds in the mucosa.
This might have compromised the seminal vesicular function.
WHO have recommended the determination of this sugar to assess the
function of these glands.
The present study measures the fructose levels in adult mice after
administration of a range of kisspeptin doses to evaluate the function of
seminal vesicles after kisspeptin exposure.

Materials and Methods

Forty (n=40), adult male albino mice ( Mus musculus) of
Swiss strain with an average weight of 505g were purchased
from the NIH, Islamabad and maintained in the animal house
facility of University throughout the experimental period.
Ten mice were housed per cage.
Standard rat diet and water were provided ad libitum.
All animal handling and subsequent sacrifice was done
according to the guidelines provided by the Institutional
Review Board which are in accordance with European Union
guidelines for use of laboratory animals.

Dosage and Treatment

Kisspeptin-10 was purchased from Calbiochem and was dissolved
in 1-ml dimethylesulphoxide (DMSO) to give a stock solution of 1
mg/ml that was diluted further with distilled water and was
administered intraperitoneally .
Animals were randomly assigned to four groups ( n = 10 in each).
Group I mice constituted control and received 0.9 % w/v physiological
saline
Group-II mice received 10 pg kisspeptin
Group-III mice received 1 ng kisspeptin
Group IV received 1g kisspeptin

as twice daily dose, after every 12 h for 12 days.

Collection of Tissue Samples

Toward the end of experiment, 3 h after the last dose of the
peptide, animals were anesthetized with sodium
pentobarbital.
Seminal vesicles were dissected out.
The excised tissues were weighed, rinsed in phosphate
buffered saline and were stored at -50C until assayed.
1 ml of distilled water was added to the tissue samples, they
were then macerated and centrifugated at 5,000 rpm, for 10
min.
Supernatant was aspirated for the quantitative determination
of seminal fructose levels.

Fructose Estimation
Quantification of seminal fructose was done
photometrically.
Fructose levels were estimated using Fructose test
kit (FertiPro N.V. Belgium).
Assays were carried out
manufacturers instructions.

according

The absorbance was measured at505 nm.

to

the

Statistical Analysis
Results were expressed as mean SEM.
The results obtained were analyzed and compared
by one-way ANOVA followed by post hoc
Tukeys adjustment using the SPSS, version16.
P<0.05 was
significant.

considered

to

be

statistically

Results
The concentration of fructose was reduced
significantly ( P<0.05) in the treated groups

Conclusion
The study reveals that chronic kisspeptin-10
administration down regulates the fructose
content of the seminal vesicles in adult male
mice.