Development of a Bicistronic Vector for the Expression of a CRISPR/Cas9-mCherry System in Fish Cell Lines.

The clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system has been widely used in animals as an efficient genome editing tool. In fish cells, the technique has been difficult to implement due to the lack of proper vectors that use active promoters to drive the expression of both small guide RNA (sgRNA) and the Cas9 (spCas9) protein within a single expression platform. Until now, fish cells have been modified using co-transfection of the mRNA of both the sgRNA and the spCas9.

Prolactin size variants during pregnancy in women with ovulatory hyperprolactinemia: characterization by isoelectric focusing and lectin affinity chromatography.

Previous studies from this laboratory have demonstrated the occurrence of important changes in PRL size heterogeneity in women with ovulatory hyperprolactinemia during gestation. A similar observation has been made, in normal women, for glycosylated PRL, which shows a progressive decrease as pregnancy progresses. In this study we decided to investigate the contribution of G-PRL on PRL heterogeneity throughout gestation in women with ovulatory hyperprolactinemia.

Reduced dopaminergic inhibition of thyrotropin release in hyperprolactinemic ovulatory women.

It has been suggested that menstrual irregularities in hyperprolactinemia are secondary to an increase in hypothalamic dopaminergic activity via a short loop positive feedback of prolactin (PRL). We have studied this question in a relatively new syndrome characterized by hyperprolactinemia without derangements of the hypothalamic-pituitary-ovarian function due to macroprolactinemia (abnormal high amounts of big-big PRL). Central dopaminergic activity was investigated by the administration of the dopamine antagonist domperidone to normal women (n = 7) and women with anovulatory (n = 6) and ovulatory hyperprolactinemia (n = 2).

[Prolactin variants in serum and amniotic fluid of women with ovulatory hyperprolactinemia].

There is enough evidence that prolactin (PRL), like many other pituitary hormones, is composed by multiple forms that differ from each other by size charge. Although they can be seen in the pituitary glands of a variety of species, their biological significance, identity and chemical nature still remain poorly understood. Recently, a new syndrome characterized by normal ovarian function in the presence of sustained hyperprolactinemia has been reported by our group.

Heterogeneity of serum prolactin throughout the menstrual cycle and pregnancy in hyperprolactinemic women with normal ovarian function.

We have demonstrated the selective secretion of high mol wt PRL series (big big PRL) in women with hyperprolactinemia and normal ovarian function. This observation suggests that big big PRL is immunologically similar, but biologically less active, than monomeric or little PRL. In this study we determined the molecular size heterogeneity of immunoreactive PRL in the serum from two ovulatory hyperprolactinemic women (subjects A and B) who had large amounts of serum big big PRL during a menstrual cycle and/or gestation.