ABSTRACT
Tubulin genes encode a series of homologous proteins used to construct microtubules which are essential for multiple cellular processes. Neural development is particularly reliant on functional microtubule structures. Tubulin genes comprise a large family of genes with very high sequence similarity between multiple family members. Human genetics has demonstrated that a large spectrum of cortical malformations are associated with de novo heterozygous mutations in tubulin genes. However, the absolute requirement for many of these genes in development and disease has not been previously tested in genetic loss of function models. Here we directly test the requirement for Tuba1a, Tubb2a and Tubb2b in the mouse by deleting each gene individually using CRISPR-Cas9 genome editing. We show that loss of Tubb2a or Tubb2b does not impair survival but does lead to relatively mild cortical malformation phenotypes. In contrast, loss of Tuba1a is perinatal lethal and leads to significant forebrain dysmorphology. We also present a novel mouse ENU allele of Tuba1a with phenotypes similar to the null allele. This demonstrates the requirements for each of the tubulin genes and levels of functional redundancy are quite different throughout the gene family. The ability of the mouse to survive in the absence of some tubulin genes known to cause disease in humans suggests future intervention strategies for these devastating tubulinopathy diseases.
Subject(s)
Malformations of Cortical Development/genetics , Neurogenesis/genetics , Sensorimotor Cortex/embryology , Tubulin/genetics , Alleles , Animals , CRISPR-Cas Systems , Disease Models, Animal , Embryo, Mammalian , Ethylnitrosourea/toxicity , Female , Gene Deletion , Gene Editing , Gene Expression Regulation, Developmental , Humans , Male , Malformations of Cortical Development/mortality , Malformations of Cortical Development/pathology , Mice , Mice, Transgenic , Microtubules/genetics , Models, Animal , Mutagenesis/drug effects , Sensorimotor Cortex/abnormalities , Species Specificity , Tubulin/metabolismABSTRACT
Clinical studies consistently report structural impairments (i.e.: ventricular enlargement, decreased volume of anterior cingulate cortex or hippocampus) and functional abnormalities including changes in regional cerebral blood flow in individuals suffering from schizophrenia, which can be evaluated by magnetic resonance imaging (MRI) techniques. The aim of this study was to assess cerebral blood perfusion in several schizophrenia-related brain regions using Arterial Spin Labelling MRI (ASL MRI, 9.4 T Bruker BioSpec 94/30USR scanner) in rats. In this study, prenatal exposure to methylazoxymethanol acetate (MAM, 22 mg/kg) at gestational day (GD) 17 and the perinatal treatment with Δ-9-tetrahydrocannabinol (THC, 5 mg/kg) from GD15 to postnatal day 9 elicited behavioral deficits consistent with schizophrenia-like phenotype, which is in agreement with the neurodevelopmental hypothesis of schizophrenia. In MAM exposed rats a significant enlargement of lateral ventricles and perfusion changes (i.e.: increased blood perfusion in the circle of Willis and sensorimotor cortex and decreased perfusion in hippocampus) were detected. On the other hand, the THC perinatally exposed rats did not show differences in the cerebral blood perfusion in any region of interest. These results suggest that although both pre/perinatal insults showed some of the schizophrenia-like deficits, these are not strictly related to distinct hemodynamic features.
Subject(s)
Dronabinol/toxicity , Methylazoxymethanol Acetate/toxicity , Neurogenesis/drug effects , Prenatal Exposure Delayed Effects/diagnostic imaging , Schizophrenia/chemically induced , Animals , Behavior Observation Techniques , Cerebrovascular Circulation/drug effects , Circle of Willis/diagnostic imaging , Circle of Willis/drug effects , Circle of Willis/embryology , Disease Models, Animal , Female , Hippocampus/blood supply , Hippocampus/diagnostic imaging , Hippocampus/drug effects , Hippocampus/embryology , Humans , Magnetic Resonance Angiography/methods , Male , Pregnancy , Prenatal Exposure Delayed Effects/chemically induced , Rats , Schizophrenia/diagnosis , Sensorimotor Cortex/blood supply , Sensorimotor Cortex/diagnostic imaging , Sensorimotor Cortex/drug effects , Sensorimotor Cortex/embryologyABSTRACT
Functional circuits of the human brain emerge and change dramatically over the second half of gestation. It is possible that variation in neural functional system connectivity in utero predicts individual differences in infant behavioral development, but this possibility has yet to be examined. The current study examines the association between fetal sensorimotor brain system functional connectivity and infant postnatal motor ability. Resting-state functional connectivity data was obtained in 96 healthy human fetuses during the second and third trimesters of pregnancy. Infant motor ability was measured 7 months after birth using the Bayley Scales of Infant Development. Increased connectivity between the emerging motor network and regions of the prefrontal cortex, temporal lobes, posterior cingulate, and supplementary motor regions was observed in infants that showed more mature motor functions. In addition, females demonstrated stronger fetal-brain to infant-behavior associations. These observations extend prior longitudinal research back into prenatal brain development and raise exciting new ideas about the advent of risk and the ontogeny of early sex differences.
