From
/09/CaCl-2006-Jana-calcium-chloride-rats.pdf:SplitDik (imported) wrote: Mon Oct 22, 2012 3:28 pm http://www.stray-afp.org/nl/wp-content/uploads/2012
The degeneration of germ cells in the CaCl2-treated testis
is possibly mediated by apoptosis. This is supported by the
appearance of some multinucleated giant cells in the
seminiferous tubules, as it is an important feature of germ
cell apoptosis [42,43]. Apoptosis of testicular germ cells by
CaCl2
treatment is further supported by the sloughing of
immature germ cells (shed cells) in the intraluminal space
[42]. Moreover, the appearance of exfoliated germ cells in
the seminiferous epithelium is also an indication that germ
cells are undergoing apoptosis, which is consistent with other
findings [43]. Germ cell detachment from the basal lamina
also indicates the induction of apoptosis in testicular tissue,
which is supported by others [42,44]. Apoptosis of germ
cells in the testis is possibly due to the low concentration of
testosterone, as testosterone is a cell survival factor for germ
cells in the testis, and withdrawal of testosterone induces
apoptosis in testicular germ cells [45]. Reduced concentration of intratesticular testosterone activates testicular caspase-3 activity, which is associated with germ cell apoptosis
[46]. Moreover, the apoptotic property of CaCl2 through the
induction of caspase-3 activity was also reported by others
[47]. It is important to note that necrosis and apoptosis can
occur simultaneously [48], and, after that, apoptotic cells
undergo secondary necrosis [49].
Another way in which testicular degeneration was noted
by CaCl2 may be through the generation of reactive oxygen
species (ROS) or their products in testicular tissue, as ROS
production in the testis results in a low level of testosterone,
which is associated with germ cell degeneration [50,51].
Calcium chloride is also an effective chemical agent for
inducing the generation of ROS in tissues [52]. This is
supported by the appearance of giant cells in seminiferous
tubules in the treated group, and these giant cells are also
actively engaged in free radical production [53]. Calcium
chloride-induced testicular oxidative stress can be assessed
by the measurement of MDA [54]. The fact that an
intratesticular injection of CaCl2 was associated with freeradical production in the testis was reflected by the high
testicular content of MDA. Free-radical production in the
testis of treated rats is supported by the infiltration of
leucocytes in the seminiferous tubules, as testicular freeradical production is closely associated with leucocyte
infiltration [55].
This elevation in the formation of free radicals in the
testes treated with CaCl2 is also supported by the diminution
in testicular GPx, SOD and GST activities, as these are
considered important scavenging enzymes against free
radicals in male gonads [56,57]. Besides these enzymatic
antioxidants, CaCl2 treatment was also associated with a low
content of testicular reduced GSH, an important nonenzymatic antioxidant in the testis [58]. The low activities of
testicular GPx, SOD and GST along with a low content of
GSH among the treated rats may be due to the low
concentrations of testosterone [50,51] or the direct effect of
CaCl2
, which is not explored properly in this study. It may be
assumed that testicular degeneration in this experiment was
due to generation of free radicals in the testis, as free radicals
are inhibitors for spermatogenesis [19] and testicular
androgenesis [59]. In addition, the reduced levels of GST
activity and GSH content in the testes would be another
cause for the degenerative necrosis of seminiferous tubules.
Glutathione S-transferase and GSH are important regulators
for proliferation and differentiation of germ cells and provide
protection to germ cells against the harmful effects of free
radicals [60]. However, Bauche et al. [61] showed that
Sertoli and peritubular cells have a high content of GSH and
a higher activity of GST. Degeneration of these cells by
CaCl2 treatment could be a possible cause for the lower
content of GSH and GST activity in the treated testis.
Rapid production of ROS results in DNA fragmentation, protein degradation that may lead to germ cell
K. Jana, P.K. Samanta / Contraception 73 (2006) 289 – 300 297apoptosis [57]. Besides this, improper functioning or
diminution in the activities of GPx, GST, SOD and content
of GSH in testis may also lead to germ cell apoptosis, as
these enzymes act as an anti-apoptotic factor [57,62–64]. In
CaCl2-treated rats, the diminution in prostato-somatic,
seminal vesiculo-somatic and epididymal somatic indices
may be due to low plasma concentration of testosterone, as
growth of these accessory sex organs is solely controlled by
testosterone [65]. Moreover, the low plasma concentration of
testosterone in CaCl2-treated rats has been further indicated
here by significant diminution in epididymal sperm count, as
sperm maturation in the epididymis is controlled by
testosterone [38]. Peritubular space in testicular sections of
CaCl2-treated rats also exhibited significant degenerative
changes along with proliferation of fibrous tissue. Such
changes may be due to the low level of plasma testosterone
[66]. Moreover, fibrosis of peritubular space in treated
animals is also consistent with previous studies on other
chemical agents [6,12]. The infiltration of leucocytes into the
tubular and peritubular zones is possibly due to the
degeneration of germ cells, which may release a large
amount of proteins that defuse out to the luminal cavity of
the seminiferous tubule and into the interstitium where they
act as antigen to promote chemotaxis of leucocytes [13].
Note I've been contacted by two people recently. One was injected on Monday and another will be today. They're logging their experiences, including video/photos, and assuming they are really doing it I should have something to post in about a month. Note the guy that did it on Monday said there was no pain at all during the injection, on the second day it felt like having "blue balls" but no serious pain, and today back to painless.