Cre-Controlled CRISPR mutagenesis provides fast and easy conditional gene inactivation in zebrafish.

Conditional gene inactivation is a powerful tool to determine gene function when constitutive mutations result in detrimental effects. The most commonly used technique to achieve conditional gene inactivation employs the Cre/loxP system and its ability to delete DNA sequences flanked by two loxP sites. However, targeting a gene with two loxP sites is time and labor consuming. Here, we show Cre-Controlled CRISPR (3C) mutagenesis to circumvent these issues. 3C relies on gRNA and Cre-dependent Cas9-GFP expression from the same transgene. Exogenous or transgenic supply of Cre results in Cas9-GFP expression and subsequent mutagenesis of the gene of interest. The recombined cells become fluorescently visible enabling their isolation and subjection to various omics techniques. Hence, 3C mutagenesis provides a valuable alternative to the production of loxP-flanked alleles. It might even enable the conditional inactivation of multiple genes simultaneously and should be applicable to other model organisms amenable to single integration transgenesis.

Lamin B1 decline underlies age-related loss of adult hippocampal neurogenesis.

Neurogenesis in the adult hippocampus declines with age, a process that has been implicated in cognitive and emotional impairments. However, the mechanisms underlying this decline have remained elusive. Here, we show that the age-dependent downregulation of lamin B1, one of the nuclear lamins in adult neural stem/progenitor cells (ANSPCs), underlies age-related alterations in adult hippocampal neurogenesis. Our results indicate that higher levels of lamin B1 in ANSPCs safeguard against premature differentiation and regulate the maintenance of ANSPCs. However, the level of lamin B1 in ANSPCs declines during aging. Precocious loss of lamin B1 in ANSPCs transiently promotes neurogenesis but eventually depletes it. Furthermore, the reduction of lamin B1 in ANSPCs recapitulates age-related anxiety-like behavior in mice. Our results indicate that the decline in lamin B1 underlies stem cell aging and impacts the homeostasis of adult neurogenesis and mood regulation.

Fifty years after: A taxonomic revision of the amphibian species from the Ecuadorian biodiversity hotspot Abra de Zamora, with description of two new Pristimantis species.

Abra de Zamora is an important biodiversity hotspot in southern Ecuador. Between 1938 and 2010, eleven species of frogs were described from here: Lynchius flavomaculatus, Gastrotheca psychrophila, Pristimantis balionotus, P. colodactylus, P. cryptomelas, P. percultus, P. versicolor, P. vidua, Telmatobius cirrhacelis, P. andinognomus, and Atelopus podocarpus. Unfortunately, many of these species were not re-encountered after their original description, and for the majority DNA samples were not available, making their phylogenetic position unknown. In this study, we assess the current state of the amphibians from Abra de Zamora, by: i. redescribing the species which were first reported from the area, by contributing genetic delimitation (for L. flavomaculatus, P. balionotus, P. colodactylus, P. percultus, and P. vidua), release call (L. flavomaculatus) and advertisement call descriptions (for P. balionotus, P. vidua and P. versicolor); ii. presenting an updated amphibian species list of Abra de Zamora, with the description of two additional Pristimantis species; iii. updating the distribution of these species, including data collected in similar montane habitats from surrounding areas; and iv. amending recommendations regarding their conservation status.

A new minute Pristimantis (Amphibia: Anura: Strabomantidae) from the Andes of southern Ecuador.

We describe a new rainfrog species (Pristimantis), from the wetland complex Oña, Nabón, Saraguro and Yacuambi, in the Andes of southern Ecuador, at altitudes ranging between 3000-3400 m a.s.l. Pristimantis tiktik sp. nov. is a small frog, displaying sexual dimorphism (the males with dorsum of various shades of gray, brown, orange or green and a whitish or pinkish yellow venter; females with brownish gray or gray dorsum and a reticulated white and black venter), with SVL ranging between 19.7-20.4 mm in females (n = 3) and 16.1-18.4 mm in males (n = 6). The skin on dorsum is tuberculated, that on venter is coarsely areolate, dorsolateral folds are absent, tympanic membrane is absent but the tympanic annulus is evident, cranial crests are absent, discs on fingers just slightly expanded, heel is lacking enlarged tubercles, inner edge of tarsus is bearing a long fold, Toe V is slightly longer than Toe III and the iris coloration is bronze with fine black reticulations. The males have a large subgular vocal sac that extends onto the chest and vocal slits but lack nuptial pads. The unique advertisement call consists of long duration series of periodically repeated clicks: "tik". Molecular analyses place the new species in the recently resurrected P. orestes group, as the sister species of the assemblage that contains P. bambu, P. mazar, P. simonbolivari and an undescribed species.